OBJECTIVE To find out the situation of the use of preventive application of antimicrobial in orthopedic perioperative patients and to evaluate the rationality of use. METHODS To make a survey of the use of antimicrobials in orthopedic perioperative patients during Apr-Jun 2004 and to evaluate the kinds,frequency,combined administration,course,effectiveness,etc. RESULTS 99.2% patients had received antimicrobials involving 14 kinds.62.8% from them used one.34.5% used two and 2.7% used three. CONCLUSIONS It is necessary to strengthen the management of preventive antimicrobial in orthopedic patients during perioperative period.

HCV NS5B, acting as a RNA dependent replication enzyme,has emerged as an attractive protein used as a target for screenig of drugs against HCV NS5B, and plays an important role in HCV replication. In the report the gene expression of NS5B in E.coli was investigated. PCR was performed to gain the gene of HCV NS5B from plasmid pBRTM/HCV 1 which contains whole sequence of HCV, and the truncated NS5B gene containing no hydrophobic domain was cloned into pGEM Teasy vector. The gene of the truncated NS5B was cut from pGEM Teasy vector and cloned into E.coli expression plasmid pET 21b, then pET 21b NS5B was transfected into E.coli cells. The protein E.coli lysates were purified and analyzed by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS PAGE) and Western blotting assay. The RdRp activity of NS5B was examined by scintillation proximity assay (SPA). The truncated NS5B gene was successfully cloned into pET 21b. The results of SDS PAGE and Western blotting assay showed: ①the molecular weight of the expressed product was about 68 000 D, ②The truncated NS5B protein was existed in media of E.coli cells, ③The activity of NS5BDCT21 His from HCV 1b amounted to 6 900 cpm (total incorporation of approximately). These findings suggest that soluble NS5B can be successfully expressed in E.coli and could secret into media.

Objective To clarify the actions of tranilast,an anti allergic drug,on chronic constriction mononeuropathy through an experiment using an animal model according to the method of Bennett et al. Methods 36 rats were divided into 2 groups: one was treated with tranilast(200mg/kg?d-1,p.o every experimental day),the other was controlled with solvent only. Chronological changes of heat evoked withdrawal latencies on a hot plate,nerve coducting velocitics, and histopathological changes were compared between the two groups from one to four weeks after loose ligation in the sciatic nerve. Results The changes of the withdrawal latency and nerve conducting velocities in the tranilast treatment group were significantly longer than those in the control. The rats without tranilast treatment showed inflammation in and around the constricted nerve bundles,axonal degeneration,phagocytes invasion and interstitial edematous changes at 7 days,numerous axonal sprouts and remyelination at 14 days,and regeneration in the nerve bundles at 28 days. In contrast,the rats treated with tranilast showed less inflammation or nerve fiber degeneration,and better regeneration than those of the control. Conclusion The actions of tranilast appear to have beneficial effects on prophylaxis of a chronic constriction mononeuropathy and on preservation of the nerve functions in rats.

Objective To select the optimum conditions of Echinococcosis ELISA kit and kit preparation by using orthogonal experiments,then to test their detection performance.Methods Taking the absorbance of the samples as the indicator,orthogonal experiments of three factors four levels were taken to optimize the conditions of hydatid disease detection ELISA kit.Through L16 (43)orthogonal experiments,the concentration of the antigen(A),the dilution multiple of the sample (B) and the dilution multiple of the enzyme labeled antigen (C) were studied.The specimen were provided by The First Affiliated Hospital of Xinjiang Medical University specimen library collected between March 2011 and June 2013,including 36 sera confirmed alveolar Echinococcus by the gold standard,56 sera confirmed Echinococcus granulosus disease by the gold standard and 72 sera as the control group(including healthy people,cirrhosis,hepatitis,etc).Detecting sensitivity and specificity were compared using F test for statistical analysis.Results Orthogonal design showed the size proportion of three factors of echinococcosis ELISA kit:antibody dilution ＞ sample dilution ＞ antigen coating concentration.The optimal preparation conditions were A1 B2 C4,that was,the concentration of antigen was 1 mg/L,sample dilution of 1∶ 100,dilution of secondary antibody of 1∶160 000.Through the 3-factor and 4-level orthogonal design using F-test analysis,the fact of the concentration of the kit antigen(A),F =1.181,P =0.393; the fact of the dilution multiple of the sample (B),F =2.544,P =0.152,the fact of enzyme labeled antigen dilution (C) F =2.544,P =0.039.The sensitivity of the kit to Echinococcus granulosus disease and alveolar Echinococcus were 83％ (30/36),and 91％ (51/56),respectively; the specificity was 91％ (29/32) and 83％ (33/40); the misdiagnosis rate of 17％ (6/36) and 9％ (5/56) ; the misdiagnosis rate 9％ (3/32) and 18％ (7/40) ; positive likelihood ratio 8.86％ (83/9) and 5.20％ (91/18) ; negative likelihood ratio of 0.18％ (17/91) and 0.11％ (9/83).Conclusion Orthogonal design is a good method that can find out optimal conditions of preparation for Echinococcosis ELISA kit and improve the test performance.

Objective:To study the effects of zinc deficiency on bone histomorphometric parameters of femoral distal diaphysis in rats. Methods:Thirty Wistar rats were randomly allocated into three groups:the zinc-deficient group(ZD), the control group(Cont),and the pair-fed group(ZP). After the rats had been fed for eight weeks, the histomorphometric and dynamic parameters of the rats were analysed using bone histomorphometric method. Results:The number, volume and connectivity of trabecular bone, and the mean trabecular plate density of ZD rats were significantly decreased, but their mean trabecular plate space was significantly increased. In addition, the data showed that ZD animals had significantly decreased trabecular osteoid surface, reduced velocity of bone formation as compared with Cont and ZP animals. The results showed that in ZD rats the mineral deposit rate was significantly slow, while the mineralization lag and osteoid maturation period were obviously prolonged. Conclusion: Zinc deficiency reduces velocity of bone formation and prolongs bone mineralization and destroys bone structure.

Objeetive To explore the value of antisense imaging of 99Tcm-labeled ASON targeting mouse double minute 2(MDM2) mRNA for the diagnosis of human prostate cancer.Methods The ASON targeting MDM2 mRNA and the mismatched oligonucleotide (ASONM) were synthesized and radiolabeled with 99Tcm using the bifunctional chelator HYNIC.The labeling efficiency and radiochemical purity were investigated.Animal models of nude mice bearing human prostate cancer LNCaP were established and divided into 3 groups with 10 mice in each group.99Tcm-HYNIC-ASON,99Tcm-HYNIC-ASONM (study groups) and 99TcmO4-(control group) were injected at the dose of 7.4 MBq through the tail vein,respectively.Tumor imaging was acquired with SPECT and the tumor-to-muscle (T/M) ratio was measured.The data was compared by one-way analysis of variance.Results The labeling efficiencies of ASON and ASONM were (65.15± 2.05) ％ and (64.93±2.18) ％,respectively.Their radiochemical purity was greater than 90％.At 1,4 and 10 h post injection,the T/M ratios of 99Tcm-HYNIC-ASON group were 3.217±0.125,3.749± 0.201 and 4.028±0.186,and those of 99Tcm-HYNIC-ASONM group were 1.579t0.128,1.715±1.140 and 1.683±0.139,and control group 2.146±0.132,1.847±0.124,1.528±0.152,respectively.The T/M ratios in control group and 99Tcm-HYNIC-ASONM group were significantly lower than those in 99Tcm-HYNICASON group at 1,4 and 10 h,respectively (F=213.37-235.41,t=3.527-4.738; all P＜0.01).The T/M ratios of 99Tcm-HYNIC-ASONM group and control group were not significantly different at 1,4 and 10 h (t=2.154,2.287 and 2.236,all P＞0.05).Conclusion The antisense probe of MDM2 can accumulate specifically in prostate cancer tissue in animal models,which might be useful as a non-invasive genetic tool for the early diagnosis of prostate cancer.

Objective:To study the optimal extraction process of Yinxiejing. Methods: An orthogonal experimental method was used with reflux time, reflux times, ethanol concentration and volume as the influencing factors, and emodin and osthole content as the indices. Results:The optimal extraction conditions for Yinxiejing were as follows:refluxing twice with 2 h every time using 8-fold 80%ethanol. Conclusion:The optimum process is stable and feasible.

Objective To investigate the effect of mouse double minute 2 (MDM2) mRNA ASON and mismatched oligonucleotide (ASONM) radiolabeled with 99Tcm on target gene expression in LNCaP cells.Methods The ASON and ASONM targeted to MDM2 mRNA were synthesized and radiolabeled by 99Tcm with the bifunctional chelator of HYNIC.The labeling efficiency,radiochemical purity,stability and molecular hybridization activity were investigated.The different concentrations of 99Tcm-HYNIC-ASON (0,100,500 nmol/L) and 99Tcm-HYNIC-ASONM (500 nmol/L) coated with lipofectamin 2000 were incubated with prostate cancer cells for 24 h,then RT-PCR and Western blot were carried out to assay the MDM2,p53 mRNA and the corresponding protein level.The variables of RT-PCR and Western blot were analyzed using one-way analysis of variance and q test.Results The labeling efficiency of ASON and ASONM were (65.15± 2.05)％ (n=5) and (64.93±2.18)％ (n=5),respectively.The radiochemical purity were both more than 90％.99Tcm-HYNIC-ASON had a good stability and could hybridize to the sense oligonucleotide (SON).The contents of MDM2 mRNA in 0,100,500 nmol/L 99Tcm-HYNIC-ASON and 500 nmol/L 99Tcm-HYNIC-ASONM groups were 0.458±0.035,0.250±0.026,0.174±0.032,0.463±0.033,respectively,and there were significant differences between each 2 groups except between 0 nmol/L 99Tcm-HYNIC-ASON and 500 nmol/L 99Tcm-HYNIC-ASONM groups (F=33.69,q =24.32-91.45,all P＜0.01).The average density of MDM2 protein in the 4 groups were 90.712±3.042,71.218±2.915,32.775±3.062,88.121±2.710,respectively (F=235.93,q=6.43-19.14,all P＜0.01; except 0 nmol/L99Tcm-HYNIC-ASON and 500 nmol/L 99Tcm-HYNIC-ASONM).The contents of p53 mRNA in the 4 groups were 0.185±0.046,0.203±0.040,0.213±0.027,0.163±0.049,respectively(F =2.18,P＞ 0.05).The average density of p53 protein was 33.865 ± 2.213,70.445±2.180,99.025±3.012,38.351±3.271,respectively (F=53.98,q =3.32-6.74,all P＜0.01 ; except 0 nmol/L 99Tcm-HYNIC-ASON and 500 nmol/L 99Tcm-HYNIC-ASONM).Conclusions The MDM2 antisense probe can accumulate in the prostate cancer cells,and specially hybridize to the MDM2 mRNA and inhibit target gene expression.This novel molecular probe has a promising potential for the diagnosis of prostate cancer at gene level.

[Summary] During recent years, increasing evidences have indicated that type 2 diabetes mellitus(T2DM) might increase the risk of certain tumors; the process might be not only related with the chronic pathologic status of T2DM such as hyperglycemia, hyperinsulinemia, insulin resistance, dyslipidemia, status of chronic inflammation but also associated with the long-term use of anti-diabetic drugs (i. e. sulfonylureas, biguanides, glitazones, dipeptidyl peptidase-4 inhibitors, glucagon-like peptitide-1 receptor agonists), as well as the use of insulin and insulin analogues. Herewith a system review was made about the recent progress in studying T2DM and tumor risk.

Objective To investigate the drug sensitivity and distribution of main pathogenic pathogens in hospitalized patients with respiratory hospital diabetes bacteria and provide reference for the prevention and treatment of diabetes mellitus in experimental infection in patients with respiratory hospital.Methods 69 cases of diabetes mellitus complicated with respiratory infection were selected, their clinical data ( laboratory, imaging and etiological examination data) were complete,and their intact specimens for testing and identificated were throat swabs, sputum smear, sputum culture, protected specimen brush sampling, pathogens culture, and drug susceptibility test of pathogenic bacteria were isolated from the statistical data of 69 patients and prognosis.Results 69 strains of pathogenic bacteria were isolated, identified and isolated from the specimens of all the patients in the study.Among them, there were a total of 86 strains of pathogenic bacteria, including gram positive bacteria (46 strains), gram negative bacteria (27 strains) and fungi (13 strains) .Staphylococcus aureus, Streptococcus pneumoniae, Staphylococcus epidermidis to teicoplanin, clindamycin, amoxicillin and levofloxacin sensitive rate is less than 50%, while to oxacillin, vancomycin, linezolid, ampicillin, imipenem, moxifloxacin susceptibility rate >50%.Pseudomonas aeruginosa, Escherichia coli, Klebsiella pneumoniae to ceftriaxone, ceftazidime, amoxicillin, amikacin drug sensitivity of less than 50% of cefoxitin, piperacillin/tazobactam, cotrimoxazole, imipenem, meropenem, netilmicin susceptibility rate >50%.69 patients were diagnosed before treatment with antiviral drugs, antibiotics, antibiotics adjusted according to the etiology and drug sensitivity test results and antiviral drugs, improved in 49 cases of patients to the hospital treatment of 14 cases, 6 cases were converted to sepsis after rescue invalid death.Conclusion The diabetic patients hospitalized in respiratory pathogens of nosocomial infection in a wide range of pathogen susceptibility differences , understand the distribution of pathogens in patients with regular and drug resistance, provide guidance for the prevention and treatment experience, reduce the infection rate of patients with respiratory infection in patients with hospital provided treatment of high pertinence has important significance.